This dissertation has been 64—6978 microfilmed exactly as received WILLIAMS, Russell Raymond, 1926- LIFE HISTORY STUDIES ON FOUR DIGENETIC TREMATODES THAT UTILIZE LYMNAEA (STAG- NICOLA) REFLEXA (SAY) AS THEIR FIRST INTERMEDIATE HOST IN A TEMPORARY POND HABITAT. \ The Ohio State University, Ph.D., 1963 ^^m f^rsity Microfilms, Inc., Ann Arbor, Michigan LIFE HISTORY STUDIES ON POUR DIGENECIC TREMATODES THAT UTILIZE LYMNAEA (STAGNICOLA) REFF.EXA (SAY) AS THEIR FIRST INTERMEDIATE HOST IN A TEMPORARY POND HABITAT DISSERTATION Presented in Partial fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Russell Raymond Williams, B. Sc., M. Sc. ■5H H H H H t The Ohio State University 1963 Approved by Adviser tent of Zoology Entomology / ACKNOWLEDGEMENTS I wish to extend ny sincere, appreciation to the many persons whose help has been invaluable in the completion of this study. For the help and guidance given me during the preparation of this thesis I am especially indebted to my adviser, Dr. Joseph N. Miller. In addition, his patience and understanding have been greatly appreciated throughout my years of graduate training. I wish to express my thanks to Dr. Allen McIntosh for the loan of specimens from the U. S. National Museum Helminthological Collection, and to Dr. Henry van der Schalie for identifying the species of lymnaeid snail used in this research. I wish also to thank the following: Dr. Robert Shields for sectioning and staining material for me as well as for his many helpful suggestions} Mr. James McGraw for helping me in the laboratory and on collecting trips; and Dr. John Crites and Dr. Wilbur Tidd for their interest, encouragement, and helpful suggestions. In addition, I wish to thank Mr. Lutz May for permitting me to work and collect at Calamus Swamp. To the many other persons who have helped either directly or indirectly, I am also greatly indebted. Not to be forgotten is ny wife, Carol, whose constant encouragement contributed to the completion of this study. ii TABLE OF CONTENTS Page ACKNOWLEDGEMENTS.............................................. ii LEST OF TABLES............................................. ▼ LIST OF ILLUSTRATIONS...................................... vi INTRODUCTION.......................................... ^. 1 MATERIALS AND METHODS...................................... h OBSERVATIONS AND RESULTS...................................... 10 The First Intermediate H o s t ............................... 10 The Trematode Parasites ............................. U The Taxonomic Positions of the Four Trematodes Encountered in this Investigation .................. 11 Diplostomum micradenum (Cort and Brackett, 1938)....... 13 Introduction. .................. 13 The Daughter Sporocyst........................... 18 The Cercaria.................................... 19 Penetration Experiments...................... 21 The Metacercaria......................... 2k Second Intermediate Hosts.................... 25 Recovery From Eye and Br a i n .............. 26 Feeding Experiments with Metacercariae......... 29 The Adult....................................... 32 Discussion...................................... 33 Protechinostoma mucronisertulatum Beaver, 19U3......... 37 Introduction.................................... 37 The Redia............................ U5 The Cercaria.................................... U5> Encystment................................ h i The Metacercaria................................ h i Excystment.................................. 1*8 Feeding Experiments with Metacercariae......... 1*9 The Adult....................................... 51 Discussion...................................... 53 Notocotylus stagnicolae Herber, 19U2.................. 55 Introduction. ............................. 55 The Redia ...................................... 59 iii TABLE OF C OUT EM'S (CONTINUED) Page The Cerceria.................................. 60 The Metacercaria..................... .. 62 Feeding Experiments withMetacercariae ........ 63 The Adnlt ............ 6U Discussion.................................... 66 Plagiorchis noblei Park, 1 9 3 6 ...................... 67 Introduction. ........................... 67 The Daughter Sporocyst......................... 72 The Cercaria.................................. 73 Penetration of SecondIntermediate Host .... 75 The Metacercaria............................... 77 Feeding Experiments with Metacercariae........ 77 The A d u l t .................................... 79 The Egg and Miracidium......... ... 81 Discussion.................................... 83 SUMMARY.................................................. 89 LITERATURE CITED......................................... 92 AUTOBIOGRAHIT........................................... 96 iv LIST OF TABLES Table Page 1 1961 Collection and Infection Data for Iymnaea (S.) reflexa.................................. “ ....... I k 2 1962 Collection and Infection Data for lynmaea (S.) reflexa.................................. - ....... 15 3 1963 Collection and Infection Data for Lymnaea (S.) reflexa.................................. “ ....... 16 U Diplostomum micradenum (Cort and Brackett, 1938) Exposure”of fish and Amphibia to Cercariae........... 22 5 Diplostomum micradenum (Cort and Brackett, 1938) Recovery of Metacercariae from Rana pipiens Tadpoles 30 Days after Exposure to $0 CercariW Each. ...... 28 6 A Two-Way Analysis of Variance for the Metacercarial Data from 19 Rana pipiens Exposed to 50 Cercariae Bach . 29 7 Diplostomum micradenum (Cort and Brackett, 1938) % Results of1 Feeding Metacercariae from Rana pipiens Tadpoles to One-Day-Old Domestic Du c k s ............... 30 8 Diplostomum micradenum (Cort and Brackett, 1938) Results of deeding Kfetacercariae from Rana pipiens Tadpoles to One-Day-Old DomesticCh i c k e n s ............ 31 9 Protechlnostoma mucronisertulatum Beaver, 19b3 Results of Feeding Metacercariae to Various Vertebrate Animals........................................... 50 10 Notocotylus stagnicolae Herber, 191*2 Results of Feeding 1- to 1;-Day-Old Metacercariae to Domestic Chickens.................................. 63 11 Plagiorchis noblei Park, 1936 Results of Feeding Metacercariae to Vertebrate Animals........................................... 78 v LIST OF ILLUSTRATIONS PIATE I . ........................................... Life History Stages of Diplostomum micradenum. Fig. 1.— Metacercariae in Retina of Eye of Rana pipiens Tadpole. PIATE II ............................................... Life History Stages of Diplostomum micradenum. Fig. 1.--Metacercariae in Vesicles of Cerebral Hemispheres of Rana pipiens Tadpole. H A T E H I ............................................... Life History Stages of Dfmlostomum micradenum. fig. 1.— Cercaria. Ventral View. Fig. 2.— Cercaria Shoving Normal Resting Position, fig. 3• —Daughter Sporocysts. Fig. U>— Eleven-Day-Old Metacercaria. Ventral View. Fig. 5>*— Thirteen-Day-Old Metacercaria. Ventral View, fig. 6 . — Adult fluke. Ventral View. PIATE IV ............................................... Life History Stages of Protechlnostoma mucronisertulatum. Fig. 1.— Cercaria. Dorsal View. Fig. 2.— Cercaria Shoving fin Fold. Lateral View. Jig* 3«— Mature Redia Containing Cercariae. fig. — Immature Redia. fig. 5.— Encysted Metacercaria. fig. 6 .— Excysted Metacercaria with Cyst Still Present, fig. 7.--Adult Fluke. Ventral View. HATE V ............................................... Life History Stages of Notoeotylus stagnicolae. fig. 1.— Daughter Redia Containing a Single Cercaria. Fig. 2.— Cercaria. Dorsal View, fig. 3.— Encysted Metacercaria. Fig. it.— Adult fluke. Ventral View, fig. 5-— filamentous Egg. LIST OF ILLUSTRATIONS (CONTINUED) PLATE V I ...................... ........................ Life History Stages of Plagiorchis noblei. Fig. 1.— Daughter Sporocyst Containing Cercariae. Fig. 2.— Cercaria. Ventral View. Fig. 3•— Stylet. Dorsal View. Jig. It.--Stylet. Lateral View. Jig. 5 .--Oral Sucker of Cercaria Showing Position of Stylet. Lateral View. Jig. 6 .--Encysted Metacercaria. Jig. 7.— Six-Day-Old Experimental Adult Jluke. Ventral View. Fig. 8 .— Adult Fluke from Natural Infection in Red-Winged Blackbird. Ventral View. PLATE VII ............................................... Life History Stages of Plagiorchis noblei. Jig. 1.— Egg from Near Metraterm of Uterus. Jig. 2.— Egg Possessing Miracidium after Jive Days of Incubation. fig. 3«~Adult Jluke. Transverse Section Through Region of Genital Pore. Fig. h-— Adult Jluke. Transverse Section Through Region of Acetabulum. Jig. 5 .--Adult Jluke. Transverse Section Through Region Posterior to Testes. INTRODUCTION This represents a stu<fy of the life histories of four species of digenetic trematodes, the asexually reproducing stages of which were found infecting the pulmonate snail, lymnaea (S.) reflexa (Say). Snails of this species were collected over a two-year period at Calamus Swamp which is a natural temporary pond near Circleville in Pickaway County, Ohio. This swamp covers approximately ten acres when full. Calamus Swamp was included in the March, 19llt, geological survey edition of the Circleville Quadrangle, and undoubtedly dates back to many years before that time. According to Stansbery (personal communication), the underlying floor of the swamp consists primarily of a clay lens which forms an impervious layer in the glacial till of the region. Water drains into the swamp only from adjoining fields. Although considered a temporary